Non-Fourier methods of spectrum analysis have enabled previously intractable multidimensional NMR experiment on biomolecules by providing high resolution spectra from data collected using nonuniform (sparse) sampling. Compared to the Fourier Transform, we know much less about the properties of these methods. In order to fully realize their potential benefits, and to understand their vulnerabilities, it is essential that we investigate their robustness, failure modes, and convergence properties. Further improvements in these methods are essential for meeting the challenges posed by larger, more complex, fleetingly stable, and dynamically disordered systems, but the lack of detailed understanding as well as the absence of robust metrics for spectral quality has hampered systematic approaches, critical comparison of competing approaches, or determination of best practices. The broad aim of this proposal is to develop robust metrics and apply them to improve the methods, and apply them to enable challenging biomolecular applications of multidimensional NMR. A domain of the protein USP7, involved in cancer, will serve as a case study. The potential impact of these developments is substantial because they will improve the power of the installed base of NMR spectrometers for biomolecular applications.

Public Health Relevance

Structural biology is the study of the shapes of biological macromolecules, and knowledge of their shapes is vital for understanding basic biology and for practical applications such as drug discovery. NMR is an important tool in structural biology as advances in technology enable the study of larger and more complex systems, and because many proteins are known to be intrinsically disordered or highly flexible, making NMR the only available method for studying such biomolecules in atomic detail. A limiting facet of NMR is its inherent insensitivity. This project involves development of modern methods of spectrum analysis ? used to convert time-domain NMR data into frequency spectra that are subsequently analyzed ? to enhance the sensitivity of NMR experiments. This improved sensitivity will extend the applicability of NMR to challenging molecules that are more complex, fleeting stable, or sparingly soluble. A domain of the protein USP7 serves as a case study.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM123249-01A1S1
Application #
9690322
Study Section
Program Officer
Smith, Ward
Project Start
2018-02-01
Project End
2022-01-31
Budget Start
2018-02-01
Budget End
2019-01-31
Support Year
1
Fiscal Year
2018
Total Cost
Indirect Cost
Name
University of Connecticut
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
022254226
City
Farmington
State
CT
Country
United States
Zip Code
Zambrello, Matthew A; Schuyler, Adam D; Maciejewski, Mark W et al. (2018) Nonuniform sampling in multidimensional NMR for improving spectral sensitivity. Methods 138-139:62-68
Delaglio, Frank; Walker, Gregory S; Farley, Kathleen A et al. (2017) Non-Uniform Sampling for All: More NMR Spectral Quality, Less Measurement Time. Am Pharm Rev 20: